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- 2025年07月10日
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- 详细信息
- 文献和实验
- 技术资料
- 供应商：
上海研卉生物科技有限公司
- 保存条件：
-20℃
- 库存：
大量
- 英文名：
LDH-Cytotoxicity Assay Kit II
- 规格：
500次分析
Kit Summary:
• Detection method- Absorbance (450 nm)
• Sample type- Cell culture (adherent and suspension)
• Species reactivity- Mammalian
• Applications- This assay provides a convenient and Wst-based non-radioactive assay for measurement of activity of lactate dehydrogenase (LDH) which is a stable enzyme normally found in the cytosol of all cells but rapidly releases into the supernatant upon damage of plasma membrane.
Features & Benefits:
• Simple one-step procedure; takes around than 1 hour
• Fast and convenient
Kit components:
• WST Substrate Mix
• LDH Assay Buffer
• Cell Lysis Solution
• Stop Solution
• LDH
Description:
Cell death or cytotoxicity is classically evaluated by the quantification of plasma membrane damage. Lactate dehydragenase (LDH) is a stable enzyme, present in all cell types, and rapidly released into the cell culture medium upon damage of the plasma membrane. LDH, therefore, is the most widely used marker in cytotoxicity study. BioVision’s LDH Cytotoxicity Assay Kit II utilizes the advanced WST reagent for a fast and more sensitive detection of LDH released from damaged cells. The assay utilizing an enzymatic coupling reaction: LDH oxidizes lactate to generate NADH, which then reacts with WST to generate yellow color. The intensity of the generated color correlates directly with the cell number lysed. Since WST is brighter, less amount of culture medium is required for the assay, and thus the background from serum and culture medium is significantly reduced. Using the assay, cells can be cultured in regular 10% serum containing medium, no reducing serum or special medium is required for the assay. In addition, since the WST is more stable, the reaction can be read multiple times, and can also be stopped at any time point during the reaction. LDH activity can be easily quantified by spectrophotometer or plate reader at OD450 nm. The kit provides all necessary reagents including LDH positive control. Assay takes less than 1 hour.
Storage Conditions:
-20°C
Shipping Conditions:
gel pack
USAGE: For Research Use Only! Not For Use in Humans.
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文献和实验1. 原理：LDH (乳酸脱氢酶） 是稳定的胞浆酶，存在所有的细胞中，当胞膜损伤时快速释放到细胞培养液中。 LDH 活性通过两个酶催化反应： LDH 氧化乳酸盐生成丙酮酸盐，然后丙酮酸盐和四唑盐 INT 反应生成甲（ formazan ）结晶。甲（ formazan ）结晶量在培养液中的增加，与裂解的细胞数增加直接相关。甲（ formazan ）结晶染料是水溶的，可以用分光光度计在 500nm 波长检测。通过检测细胞培养上清中
基因敲除模型：NLRP3−/−、GSDMD−/−、caspase-1/11−/−小鼠、RACK1ΔMΦ。 4、检测内容：LDH（乳酸脱氢酶）释放实验检测细胞毒性、ELISA 检测细胞因子分泌、WB 检测、免疫沉淀（IP）、pull down、质谱、EST12 晶体结构、共聚焦显微镜、小鼠结核感染模型等。 研究结果： 1、EST12（Rv1579c）诱导 GSDMD 介导的巨噬细胞焦亡 表达纯化 40 种 RD 区编码蛋白，通过 LDH 释放实验，发现 EST12（Rv1579c）具有明显的巨噬细胞毒性
细胞毒检测技术包括补体依赖细胞毒试验、NK细胞介导的细胞毒试验和特异性CTL活性检测，常用的实验技术有后两种。NK细胞（natural killer cell）是在天然免疫系统发挥主要作用的效应细胞，可直接杀伤病毒感染的靶细胞和某些肿瘤细胞。而细胞毒性T淋巴细胞（cytotoxic T lymphocytes，CTL）则是特异免疫系统发挥特异杀伤作用的效应细胞，可特异杀伤病毒感染细胞和肿瘤细胞。本节介绍乳酸脱氢酶(LDH)释放法检测NK细胞活性、Calcein释放实验检测CTL活性
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